ImmoMix™

Ordering

Description

ImmoMix™ is a complete ready-to-use high yield heat-activated 2x reaction-mix, which simply requires the user to add water, template and primers, and then pre-heat to 95°C for 10 minutes to carry out successful PCR assays.

Product Highlights

Convenient - pre-mixed, pre-optimized 2x solution

Ready to use format - reduces risk of contamination

Fast set up - decreased time compared to traditional methods

Reproducible results - time after time

Product Description

ImmoMix™ is a complete ready-to-use high yield, heat-activated 2x reaction-mix, which simply requires the user to add water, template and primers, and then pre-heat to 95°C for 10 minutes to successfully carry out PCR assays. The 10 minute activation step eliminates the presence of non-specifics such as primer-dimers and mis-primed products, since the enzyme is inactive at initial low temperatures.

ImmoMix is based on IMMOLASE™ DNA Polymerase, which leaves an ´A´ overhang, and has been optimized for a wide variety of templates. Additional MgCl2 solution is included should any fine adjustments be required.

ImmoMix reduces the time needed to set up reactions, thereby reducing the risk of contamination. Greater reproducibility is ensured by reducing the number of pipetting steps that can lead to pipetting errors.

Applications

Ultra-high specificity for multiplex reactions

Products suitable for TA cloning

Fig. 1 Higher yields from ImmoMix

To illustrate the higher yield of ImmoMix, an 800 bp fragment of the β-actin gene was amplified from human genomic DNA, using 100 ng (lane 1) followed by 5-fold serial dilutions (lanes 2-6). This was compared to three other suppliers of chemical hot-start polymerases, using the manufacturers' recommended protocol. The results illustrate the higher yield using ImmoMix than the other suppliers of hot-start polymerases. HyperLadder 50bp (M).

Specification

Components

Reagent

500 Reactions

ImmoMix

10 x 1.25 mL

50 mM MgCl2 Solution

1.2 mL

Volume

BIO-25020: 500 x 50 µL Reactions: 10 x 1.25 mL

Concentration

2x

Storage & Stability

All components should be stored at -20°C upon receipt for optimum stability. Repeated freeze/thaw cycles should be avoided. When stored under the recommended conditions and handled correctly, full activity of the reagents is retained until the expiry date on the outer box label.

Components may also be stored at +4°C if required, although storage at -20°C is recommended. When stored at +4°C, reagents will remain stable for a period of two weeks from date of receipt.

Shipping conditions

On Dry Ice or Blue Ice.

FAQs

Bioline's mixes contain magnesium chloride at the following concentrations (please see table below). An additional tube of 50 mM MgCl2 solution is supplied with each mix to facilitate further optimization if required.

PCR can be a challenging technique, with various parameters to optimize to achieve the best results. If you are having problems, these could be easily resolved by addressing a few issues. Please see our PCR troubleshooting guide for suggestions and help with your specific problems.

These terms refer to parameters to be considered when performing PCR and are important features in selecting the correct enzyme for your needs. Understanding what they mean is therefore crucial:
Yield: The amount of DNA produced in a PCR reaction.
Fidelity: The accuracy of the enzyme at incorporating the correct dNTP to the elongating DNA strand.
Processivity: The length of time a polymerase is associated with the template and therefore the size of fragment which can be amplified.
Specificity: A measure of the unwanted by-products generated in a reaction.

During PCR setup at room temperature, standard polymerases will have some activity. When using a hot-start polymerase, this enzyme will have no activity at room temperature, thus reducing the risk of unspecific products in the reaction due to these mis-primed oligonucleotides. This feature makes this type of polymerase ideally suited to high-throughput applications, where the reactions may be left sitting at room temperature for prolonged periods of time.

In nature, the ability of a DNA polymerase to correct misincorporations of nucleotides in the DNA strand being elongated is often crucial to the survival of the host organism. This ability is termed "proofreading activity" and occurs in the 3' to 5' direction. This activity also leads to the polymerase removing unpaired nucleotides overhanging at the 3'end (A-overhangs), creating blunt ends.

All Bioline polymerases are available in the convenient format of a 2x master mix, containing all the reagents and additives necessary to perform successful PCR, with the exception of template, primers and water.
This formulation not only provides a convenient and hassle-free PCR setup, but also significantly reduces the chances of human error, inaccuracies and contamination by reducing the pipetting steps required.

Whilst the exact composition of the mixes is proprietary information, all mixes are made up of reaction buffer, magnesium, dNTPs, polymerase and additives, designed to keep the polymerase stable in the mix, as well as provide the optimal conditions for PCR.

If possible, we would recommend an additional cleanup of affected samples, for instance, customers had very good experiences with the clean-up of samples containing humic acids with SureClean Plus. Alternatively decreasing the template concentration will help to minimize the amount of inhibitors. The amount of primer can be increased, but do not exceed the suggested limits.